Functional characterization of a naturally occurring trans-splicing intein from Synechococcus elongatus in a mammalian cell system
Functional characterization of a naturally occurring trans-splicing intein from Synechococcus elongatus in a mammalian cell system
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DOI：
10.1016/j.ab.2010.08.018
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发表时间：
2010-12-15
影响因子：
2.9
通讯作者：
Zhou, Naiming
中科院分区：
文献类型：
--
作者：
Chen, Linjie;Zhang, Yaping;Zhou, Naiming
我们已经克隆并鉴定了一个自然产生的能够在蓝藻细长聚球藻(Se！)中反式剪接的微小DNAE内切蛋白。DNAE INTERIN)。谢了！DNAE内含素与聚球藻同源。PCC6803(SSP)DNAE内含子和发菜DNAE内含子(NPU)DNAE内含子的蛋白质序列同源性分别为60%和61%。我们的结果表明，分裂的报告，分裂的Renilla荧光素酶(Rluc)和增强型绿色荧光蛋白(EGFP)，可以在哺乳动物细胞中通过Sel Dna内含子介导的反式剪接来重组。基于Sel DNAE内含子介导的Split Rluc重组，建立了一种模拟人类免疫缺陷病毒(HIV)进入的细胞-细胞融合试验，并验证了该方法可用于筛选和药理学表征潜在的HIV进入靶向抑制剂。(C)2010 Elsevier Inc.保留所有权利。
We have cloned and characterized a naturally occurring split mini-DnaE intein capable of protein trans-splicing in the cyanobacterium Synechococcus elongatus (Se! DnaE intein). Se! DnaE intein is homologous to Synechocystis sp. PCC6803 (Ssp) DnaE intein and Nostoc punctiforme (Npu) DnaE intein, with a protein sequence identity of 60% for the N-terminal part of intein and 61% for the C-terminal part of intein. Our results demonstrate that the split reporters, split Renilla luciferase (Rluc) and enhanced green fluorescent protein (EGFP), can be reconstituted via Sel DnaE intein-mediated trans-splicing in mammalian cells. Based on Sel DnaE intein-mediated reconstitution of split Rluc, a human immunodeficiency virus (HIV) entry-mimicking cell-cell fusion assay was developed and validated as a useful assay for screening and pharmacologically characterizing potential HIV entry-targeting inhibitors. (C) 2010 Elsevier Inc. All rights reserved.